LOCUS Exported 8459 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS pcDNA3-TLR1-YFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8459) TITLE TLR Signaling Plasmids REFERENCE 2 (bases 1 to 8459) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8459 /organism="synthetic DNA construct" /mol_type="other DNA" primer_bind complement(33..52) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" enhancer 224..603 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 604..807 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 758..778 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 852..871 /label=T7 /note="T7 promoter, forward primer" promoter 852..870 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 3266..3985 /codon_start=1 /product="enhanced YFP" /label=EYFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTFGYGLKCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(3311..3332) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3572..3591) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3919..3940 /label=EGFP-C /note="EGFP, forward primer" promoter complement(4001..4019) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(4002..4019) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(4039..4056) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 4045..4269 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 4315..4743 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4402..4421) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4612..4633 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4752..4772) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 4757..5086 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 4937..5072 /label=SV40 ori /note="SV40 origin of replication" primer_bind 4999..5018 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 5153..5947 /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /label=NeoR/KanR /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" primer_bind complement(5207..5226) /label=Neo-R /note="Neomycin resistance gene, reverse primer" primer_bind 5817..5836 /label=Neo-F /note="Neomycin resistance gene, forward primer" polyA_signal 6121..6242 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6158..6177) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 6212..6231 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(6291..6307) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(6291..6307) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(6304..6326) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 6315..6331 /label=lac operator /bound_moiety="lac repressor encoded by lacI" /note="The lac repressor binds to the lac operator to inhibit transcription in E. coli. This inhibition can be relieved by adding lactose or isopropyl-beta-D-thiogalactopyranoside (IPTG)." promoter complement(6339..6369) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6384..6405 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(6522..6539) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(6693..7281) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(6773..6792) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(7452..8312) /codon_start=1 /gene="bla" /product="beta-lactamase" /label=AmpR /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" primer_bind 8075..8094 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8313..8417) /gene="bla" /label=AmpR promoter ORIGIN 1 agatctcccg atcccctatg gtgcactctc agtacaatct gctctgatgc cgcatagtta 61 agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt 121 taagctacaa caaggcaagg cttgaccgac aattgcatga agaatctgct tagggttagg 181 cgttttgcgc tgcttcgcga tgtacgggcc agatatacgc gttgacattg attattgact 241 agttattaat agtaatcaat tacggggtca ttagttcata gcccatatat ggagttccgc 301 gttacataac ttacggtaaa tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg 361 acgtcaataa tgacgtatgt tcccatagta acgccaatag ggactttcca ttgacgtcaa 421 tgggtggagt atttacggta aactgcccac ttggcagtac atcaagtgta tcatatgcca 481 agtacgcccc ctattgacgt caatgacggt aaatggcccg cctggcatta tgcccagtac 541 atgaccttat gggactttcc tacttggcag tacatctacg tattagtcat cgctattacc 601 atggtgatgc ggttttggca gtacatcaat gggcgtggat agcggtttga ctcacgggga 661 tttccaagtc tccaccccat tgacgtcaat gggagtttgt tttggcacca aaatcaacgg 721 gactttccaa aatgtcgtaa caactccgcc ccattgacgc aaatgggcgg taggcgtgta 781 cggtgggagg tctatataag cagagctctc tggctaacta gagaacccac tgcttactgg 841 cttatcgaaa ttaatacgac tcactatagg gagacccaag cttggtaccg agctcggatc 901 tatgactagc atcttccatt ttgccattat cttcatgtta atacttcaga tcagaataca 961 attatctgaa gaaagtgaat ttttagttga taggtcaaaa aacggtctca tccacgttcc 1021 taaagaccta tcccagaaaa caacaatctt aaatatatcg caaaattata tatctgagct 1081 ttggacttct gacatcttat cactgtcaaa actgaggatt ttgataattt ctcataatag 1141 aatccagtat cttgatatca gtgttttcaa attcaaccag gaattggaat acttggattt 1201 gtcccacaac aagttggtga agatttcttg ccaccctact gtgaacctca agcacttgga 1261 cctgtcattt aatgcatttg atgccctgcc tatatgcaaa gagtttggca atatgtctca 1321 actaaaattt ctggggttga gcaccacaca cttagaaaaa tctagtgtac tgccaattgc 1381 tcatttgaat atcagcaagg tcttgctggt cttaggagag acttatgggg aaaaagaaga 1441 ccctgagggc cttcaagact ttaacactga gagtctgcac attgtgttcc ccacaaacaa 1501 agaattccat tttattttgg atgtgtcagt caagactgta gcaaatctgg aactatctaa 1561 tatcaaatgt gtgctagaag ataacaaatg ttcttacttc ctaagtattc tggcgaaact 1621 tcaaacaaat ccaaagttat caaatcttac cttaaacaac attgaaacaa cttggaattc 1681 tttcattagg atcctccagc tggtttggca tacaactgta tggtatttct caatttcaaa 1741 cgtgaagcta cagggtcagc tggacttcag agattttgat tattctggca cttccttgaa 1801 ggccttgtct atacaccaag ttgtcagcga tgtgttcggt tttccgcaaa gttatatcta 1861 tgaaatcttt tcgaatatga acatcaaaaa tttcacagtg tctggtacac gcatggtcca 1921 catgctttgc ccatccaaaa ttagcccgtt cctgcatttg gatttttcca ataatctctt 1981 aacagacacg gtttttgaaa attgtgggca ccttactgag ttggagacac ttattttaca 2041 aatgaaccaa ttaaaagaac tttcaaaaat agctgaaatg actacacaga tgaagtctct 2101 gcaacaattg gatattagcc agaattctgt aagctatgat gaaaagaaag gagactgttc 2161 ttggactaaa agtttattaa gtttaaatat gtcttcaaat atatttactg acactatttt 2221 cagatgttta cctcccagga tcaaggtact tgatcttcac agcaataaaa taaagagcat 2281 tcctaaacaa gtcgtaaaac tggaagcttt gcaagaactc aatgttgctt tcaattcttt 2341 aactgacctt cctggatgtg gcagctttag cggcctttct gtattgatca ttgatcacaa 2401 ttcagtttcc cacccatcgg ctgatttctt ccagagctgc cagaagatga ggtcaataaa 2461 agcaggggac aatccattcc aatgtacctg tgagctagga gaatttgtca aaaatataga 2521 ccaagtatca agtgaagtgt tagagggctg gcctgattct tataagtgtg actacccgga 2581 aagttataga ggaaccctac taaaggactt tcacatgtct gaattatcct gcaacataac 2641 tctgctgatc gtcaccatcg ttgccaccat gctggtgttg gctgtgactg tgacctccct 2701 ctgcagctac ttggatctgc cctggtatct caggatggtg tgccagtgga cccagacccg 2761 gcgcagggcc aggaacatac ccttagaaga actccaaaga aatctccagt ttcatgcatt 2821 tatttcatat agtgggcacg attctttctg ggtgaagaat gaattattgc caaacctaga 2881 gaaagaaggt atgcagattt gccttcatga gagaaacttt gttcctggca agagcattgt 2941 ggaaaatatc atcacctgca ttgagaagag ttacaagtcc atctttgttt tgtctcccaa 3001 ctttgtccag agtgaatggt gccattatga actctacttt gcccatcaca atctctttca 3061 tgaaggatct aatagcttaa tcctgatctt gctggaaccc attccgcagt actccattcc 3121 tagcagttat cacaagctca aaagtctcat ggccaggagg acttatttgg aatggcccaa 3181 ggaaaagagc aaacgtggcc ttttttgggc taacttaagg gcagccatta atattaagct 3241 gacagagcaa gcaaagaaac tcgagatggt gagcaagggc gaggagctgt tcaccggggt 3301 ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca gcgtgtccgg 3361 cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct gcaccaccgg 3421 caagctgccc gtgccctggc ccaccctcgt gaccaccttc ggctacggcc tgaagtgctt 3481 cgcccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca tgcccgaagg 3541 ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga cccgcgccga 3601 ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa 3661 ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc acaacgtcta 3721 tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc gccacaacat 3781 cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca tcggcgacgg 3841 ccccgtgctg ctgcccgaca accactacct gagctaccag tccgccctga gcaaagaccc 3901 caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg ggatcactct 3961 cggcatggac gagctgtaca agtaatctag agggccctat tctatagtgt cacctaaatg 4021 ctagagctcg ctgatcagcc tcgactgtgc cttctagttg ccagccatct gttgtttgcc 4081 cctcccccgt gccttccttg accctggaag gtgccactcc cactgtcctt tcctaataaa 4141 atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc tattctgggg ggtggggtgg 4201 ggcaggacag caagggggag gattgggaag acaatagcag gcatgctggg gatgcggtgg 4261 gctctatggc ttctgaggcg gaaagaacca gctggggctc tagggggtat ccccacgcgc 4321 cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg accgctacac 4381 ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc gccacgttcg 4441 ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga tttagtgctt 4501 tacggcacct cgaccccaaa aaacttgatt agggtgatgg ttcacgtagt gggccatcgc 4561 cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat agtggactct 4621 tgttccaaac tggaacaaca ctcaacccta tctcggtcta ttcttttgat ttataaggga 4681 ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga 4741 attaattctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg 4801 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg 4861 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc 4921 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca 4981 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt 5041 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc 5101 ttgtatatcc attttcggat ctgatcaaga gacaggatga ggatcgtttc gcatgattga 5161 acaagatgga ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga 5221 ctgggcacaa cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg 5281 gcgcccggtt ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaggacga 5341 ggcagcgcgg ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt 5401 tgtcactgaa gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct 5461 gtcatctcac cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct 5521 gcatacgctt gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg 5581 agcacgtact cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca 5641 ggggctcgcg ccagccgaac tgttcgccag gctcaaggcg cgcatgcccg acggcgagga 5701 tctcgtcgtg acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt 5761 ttctggattc atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt 5821 ggctacccgt gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct 5881 ttacggtatc gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt 5941 cttctgagcg ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca 6001 cgagatttcg attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg 6061 gacgccggct ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc 6121 aacttgttta ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca 6181 aataaagcat ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct 6241 tatcatgtct gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg 6301 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata 6361 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca 6421 ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc 6481 gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg 6541 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta 6601 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc 6661 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag 6721 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac 6781 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc 6841 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt 6901 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc 6961 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga 7021 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta 7081 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta 7141 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga 7201 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg 7261 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag 7321 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc 7381 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact 7441 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt 7501 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta 7561 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta 7621 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc 7681 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat 7741 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt 7801 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg 7861 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca 7921 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta 7981 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg 8041 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact 8101 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg 8161 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt 8221 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga 8281 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc 8341 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa 8401 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggg //